Zhao Yong-Qiang, Feng Hui-Wei, Jia Tao, Chen Xue-Mei, Zhang Hui, Xu An-Ting, Zhang Hai-Ling, Fan Xian-Liang
Department of Otolaryngology and Head and Neck Surgery, the Second Hospital of Shandong University, Jinan, China E-mail :
Asian Pac J Cancer Prev. 2014;15(12):4919-23. doi: 10.7314/apjcp.2014.15.12.4919.
To investigate the effect of celecoxib on telomerase activity and apoptosis in a human laryngeal squamous carcinoma cell line (Hep-2 cells).
The growth inhibition rate of Hep-2 cells in vitro was measured by MTT assay, and apoptosis by TUNEL assay and flow cytometry (FCM). The TRAP-ELISA method was used to determine telomerase activity in Hep-2 cells. The mRNA expression of human telomerase RNA component(hTR), human telomerase reverse transcriptase (hTERT) and human telomerase-associated protein(hTEP1) was determined by RT-PCR assay. Expression of Bax and Bcl-2 proteins was assessed by Western blotting.
Celecoxib can inhibit proliferation and induce apoptosis in a dose- and time-dependent manner, repress telomerase activity, decrease hTERT mRNA and Bcl-2 protein expression and increase Bax protein expression, PGE2 had no effect on telomerase.
Celecoxib had the antiproliferative and pro-apoptotic effect in Hep-2 cells. Apoptosis was accompanied by a decrease in telomerase activity which was directly correlated with hTERT mRNA and up-regulation of Bax/Bcl-2. Bcl-2 may thus play an important role in telomerase activity as well as apoptosis.
研究塞来昔布对人喉鳞状癌细胞系（Hep-2细胞）端粒酶活性及凋亡的影响。
采用MTT法检测Hep-2细胞的体外生长抑制率，采用TUNEL法和流式细胞术（FCM）检测细胞凋亡。采用TRAP-ELISA法测定Hep-2细胞的端粒酶活性。采用RT-PCR法检测人端粒酶RNA组分（hTR）、人端粒酶逆转录酶（hTERT）及人端粒酶相关蛋白（hTEP1）的mRNA表达。采用蛋白质印迹法检测Bax和Bcl-2蛋白的表达。
塞来昔布能以剂量和时间依赖性方式抑制增殖并诱导凋亡，抑制端粒酶活性，降低hTERT mRNA和Bcl-2蛋白表达，增加Bax蛋白表达，PGE2对端粒酶无影响。
塞来昔布对Hep-2细胞具有抗增殖和促凋亡作用。凋亡伴随着端粒酶活性降低，这与hTERT mRNA及Bax/Bcl-2上调直接相关。因此，Bcl-2可能在端粒酶活性及凋亡中起重要作用。
